Background/Aims: Cholangiocarcinoma (CCA) is a primary malignant neoplasm with an extremely poor prognosis. While combined chemoradiotherapy has been demonstrated to delay CCA progression to a certain extent, the absence of specific molecular biomarkers or targets significantly hinders the diagnosis and treatment of CCA. Methods: Through cross-analysis of proteomics and ADMA modificationomics, we identified DDX1 overexpressed in CCA with elevated R602-ADMA modifications. HPLC-MS/MS identified PRMT1 as the methyltransferase and USP10 as the deubiquitinating enzyme for DDX1. Immunofluorescence and nuclear-cytoplasmic partitioning experiments confirmed DDX1’s nuclear localization. GO and KEGG analyses clarify the biological functions of DDX1 in response to hypoxia. RNA-seq transcriptomics analyzed key pathways influenced by DDX1. A hydrodynamic in situ CCA mouse model was established to validate the chemopreventive effects of the PRMT1-specific inhibitor GSK715 on CCA development. Results: DDX1 promotes CCA progression both in vivo and in vitro and can be inhibited by GSK715. Mechanistically, PRMT1 mediates ADMA modification at position R602 of DDX1. This modification promotes DDX1 nuclear localization by recruiting USP10 to deubiquitinate DDX1, while simultaneously inhibiting PRMT1 degradation. DDX1 promotes the transcription of PRMT1 and USP10 by binding to the mRNA 3’UTR region, establishing a positive feedback regulatory pathway. This mechanism promotes the occurrence and development of CCA and can serve as a target for the inhibitor GSK715 to suppress CCA progression. Conclusions Our study identified DDX1-R602-ADMA modification as a novel ADMA modification in CCA. It further confirmed its pivotal role in CCA progression. Targeting the USP10-PRMT1-DDX1 axis may represent a significant therapeutic approach for CCA.

Objective:To establish a mouse model of spinal cord injury(SCI),and to investigate the effect of electroacupuncture(EA)intervention on cell apoptosis after acute SCI and its mechanism.Methods:Female C57BL/6 mice were used to establish a model of SCI,and after successful modeling,the mice were randomly divided into SCI group,EA group,and Rosiglitazone group(R group);a sham-operation group(Sham group)was also established.After successful modeling,the mice in the EA group were given EA at bilat-eral Jiaji points and Zusanli once a day for 14 days,those in the R ture,and the number of surviving cells.The EA group and the R group had a significant reduction in the expression of caspase-3 and significant increases in the expression of PPARγ and CD36,and the EA group had significant reductions in the expression of Hba-a1 and Hbb-bt.In addition,RNA-Seq and TMT/iTRAQ techniques,significant analysis,Venn analysis,and dual-omics analysis identi-fied Hba-a1 and Hbb-bt as the target genes of EA.The KEGG pathway enrichment analysis showed that EA had a significant effect on the PPAR signaling pathway.Conclusion:By regulating the PPARγ-CD36 signaling pathway,EA can promote the clearance of Hba-a1 and Hbb-bt after SCI,reduce the expression level of caspase-3,alleviate cell apoptosis,and facilitate the recovery of spinal cord nerve function.

Objective:To explore the molecular mechanism of TCM compound Shengxue Tongbian Granules in improving intestinal injury in septic rats through bioinformatics and experimental validation methods.Methods:The GSE131761 gene set was processed by bioinformatics to screen differential genes, then weighted gene co-expression network analysis (WGCNA) was applied to screen modular genes. The intersection of modular genes and differential genes was taken, and finally, the least absolute shrinkage and selection operator (LASSO) technique was applied to further obtain the key targets of sepsis, which was validated by experiments. Totally 72 SD rats were divided into sham-operation group, model group, dexamethasone group (0.15 mg/kg), Shengxue Tongbian Granules low- (0.3 g/kg), medium- (0.6 g/kg), and high-dosage (1.2 g/kg) groups, with 12 rats in each group. Corresponding drug interventions were administered to each treatment group before and 12 hours after modeling. The sham-operation group and the model group were gavaged daily with equal amounts of saline. Samples were collected after 24 hours. HE staining was used to detect the pathological morphology of intestinal tissues in each group of rats; ELISA was used to detect the levels of TNF-α, diamine oxidase (DAO), IL-6, IL-10, and myeloperoxidase (MPO) in rat serum. Immunohistochemistry was used to detect the protein expressions of MPO and neutrophil elastase (NE/LANE) in intestinal tissue, and Western blot was used to detect the protein expression of peptidyl arginine deaminase (PAD4) in intestinal tissue.Results:Seven final key genes related to sepsis were selected, namely ANXA3, CYP1B1, FCAR, LILRA5, PADI4, NOV, and S100A12. Experimental results showed that drug administration alleviated intestinal injury; compared with the model group, the levels of TNF-α, IL-6, MPO, and DAO decreased in the Shengxue Tongbian Granules high-dosage group ( P<0.05), the levels of ELANE and MPO were reduced in Shengxue Tongbian Granules low-, medium-, and high-dosage groups ( P<0.05), and PAD4 expression was reduced in the Shengxue Tongbian Granules high-dosage group ( P<0.05). Conclusion:Shengxue Tongbian Granules can improve the intestinal injury of septic rats, and the mechanism may be related to the inhibition of PAD4-mediated formation of NETs and the improvement of inflammatory response.

Objective To explore the protective effect and mechanism of Xinkang Granules-containing serum in adriamycin-induced injury of cardiomyocytes H9C2 based on cGAS-STING signaling axis.Methods Adriamycin was used to induce the H9C2 cells injury model.The cells were divided into normal group,model group,Xinkang Granules group and inhibitor group.After 24 hours of intervention,the CCK-8 method was used to detect cell survival rate,the DCFH-DA fluorescent probe was used to detect the content of cell reactive oxygen species(ROS),cell apoptosis rate was detected by flow cytometry,ELISA was used to detect the content of tumor necrosis factor-α(TNF-α)in cell supernatant,colorimetry was used to detect lactate dehydrogenase(LDH)in cells,RT-qPCR was used to detect the expression of mitochondrial transcription factor A(TFAM),cyclic guanosine-adenylate synthetase(cGAS),stimulator of interferon genes(STING)and TNF-α mRNA,Western blot and immunofluorescence were used to detect the protein expressions of cGAS and STING.Results Compared with the normal group,cell survival rate in the model group was significantly reduced(P<0.01),the ROS content was significantly increased(P<0.01),the apoptosis rate significantly increased(P<0.01),the content of TNF-α in the supernatant significantly increased(P<0.01),the activity of LDH significantly increased(P<0.01),the expression of TFAM mRNA significantly decreased(P<0.01),and the expressions of TNF-α,cGAS,STING mRNA and the protein expression of cGAS and STING significantly increased(P<0.01).Compared with the model group,cell survival rate in Xinkang Granules group and inhibitor group significantly increased(P<0.01),the ROS content significantly decreased(P<0.01),the apoptosis rate significantly decreased(P<0.01),the content of TNF-α in supernatant significantly decreased(P<0.01),the activity of LDH significantly decreased(P<0.01),the expression of TFAM mRNA significantly increased(P<0.01),and the expressions of TNF-α,cGAS,STING mRNA and the protein expressions of cGAS and STING significantly decreased(P<0.05,P<0.01).Conclusion Xinkang Granules have a protective effect on adriamycin-induced H9C2 cardiomyocytes,which may be related to the inhibition of cGAS/STING axis activation and the secretion of inflammatory factors.

Intervention in chronically activated microglia-mediated neuroinflammation is a novel approach to treat Alzheimer's disease (AD). The low permeability of the blood‒brain barrier (BBB) and non-selective distribution in the brain severely restrict AD drugs' disease-modifying efficacy. Here, an immunosuppressant TREM2-lowing antisense oligonucleotides (ASOs) and resveratrol co-loaded cationic liposome is developed as an immune reprogramming nanomodulator modified by acid-cleavable BBB-targeting peptide and microglia-targeting peptide (Res@TcMNP/ASO) for AD management. Res@TcMNP/ASO can enter brain endothelial cells via D-T7 peptides. Then D-T7 undergoes an acid-responsive cleavage, facilitating the escape of Res@MNP/ASO from endo/lysosomes to cross the BBB. The detached Res@MNP/ASO specifically targets M1-phenotype microglia via exposed MG1 peptides to prompt the simultaneous delivery of two drugs into activated microglia. This nanomodulator can not only restore the immune function of microglia through TREM2-lowing ASO but also mitigate the immune stimulation to microglia caused by reactive oxygen species (ROS) through resveratrol, thereby synergistically inhibiting the chronic activation of microglia to alleviate neuroinflammation in AD. Our results indicate that this combination treatment can achieve significant behavioral and cognitive improvements in late APP/PS1 mice.

Objective:To establish a high performance liquid chromatography-evaporative light scattering detector(HPLC-ELSD)technique for simultaneous determining the content of excipients sucrose and mannitol in meningo-coccal polysaccharide vaccine.Methods:Using NanoChrom Sugar-10Ca analytical column(300 mm × 7.8 mm)and HPLC system(Agilent 1260).With purified water as the mobile phase,a flow rate of 0.5 mL·min-1,column temperature was 80 ℃,and the injection volume was 50 μL.The evaporative light detector was based on nitrogen.The carrier gas flow rate is 3.2 L·min-1,the temperature of drift tube was 1 10 ℃,the gain value was 1,and the impactor was"mode 1".This assay was subsequently validated for its system suitability,specificity,repeatability,intermediate precision and linearity,and accuracy.The established method was used to assay the contents of the sucrose and mannitol in four batches of meningococcal polysaccharide vaccine.Results:The estab-lished HPLC-ELSD method showed good systemic suitability.Specificity validation showed that there was no inter-ference peak in the blank solvent;the separation of the target peaks between sucrose and mannitol was＞2.0.The relative standard deviations(RSD)value of peak area of sucrose and mannitol in six tests were 0.44％and 0.38％,respectively.RSD of intermediate precision of both sucrose and mannitol were lower than 2.00％,indica-ting that the precision of high performance liquid chromatography instrument was well.The linear range of two excipients were 12.5-150.0 μg·mL-1(R2＞0.99,respectively).The recovery rate of sucrose and mannitol were 95.74％-99.33％,94.37％-98.85％,respectively.There was no significant difference in the contents of sucrose and mannitol in 4 batches of meningococcal polysaccharide vaccine.Conclusion:The HPLC-ELSD method showed good specificity,precision,linearity and accuracy,and the test results were stable and reliable,so that it is suitable for simultaneous determination of sucrose and mannitol contents of meningococcal polysaccharide injections.

Purpose To analyze the ultrasonic manifestations of extrarenal extracranial malignant rhabdoid tumor(EERT)in children to enhance the awareness of its early diagnosis.Materials and Methods Clinical data of 13 children with EERT diagnosed by pathology from the Children's Hospital of Chongqing Medical University were collected retrospectively from January 2014 to March 2024.The ultrasonic manifestations of these cases were summarized and compared with their pathological results.Results Among the 13 children with EERT,10 cases had tumors located in soft tissues of the extremities,and 3 cases had tumors located in the abdomen.The tumors showed irregular shapes and indistinct borders on ultrasonic images,and the average of the maximum diameters of the tumors measured by ultrasound was(10.43±2.57)cm.The interior of the tumors mostly appeared as heterogeneous hypoechoic lesions;9 of the tumors were accompanied by small patchy regions of anechoic areas(cystic degeneration or liquefaction);9 of the tumors were accompanied by strong echoes(calcification);12 of the tumors had relatively abundant blood supply.Conclusion Ultrasonic images of EERT in children have certain characteristics.Mastering the ultrasonic manifestations could assist in the clinical diagnosis and follow-up of this disease.

Allergic rhinitis(AR)is the most common airway allergic diseases.Basophils are the classical effector cells in allergy,including AR:Both IgE and non-IgE mediated methods can activate basophils during allergic reactions and induce basophils to release various inflammatory mediators,which is accompanied by changes in the expression of multiple membrane proteins,and ulti-mately leading to the clinical manifestations of allergic diseases.IL-18 may be involved in the pathogenesis of AR by directly inducing basophil activation and inflammation response via binding to its receptor IL-18Rα.Nevertheless,the binding of IL-37b to IL-18Rα initiates the downstream anti-inflammatory signals,thus inhibiting inflammation reaction in AR.Therefore,understanding the role and mechanism of IL-18 and IL-37b in the activation of blood basophils of AR patients is of great significance for the pathogenesis,treat-ment of AR and the development of related biological agents.

Objective:To investigate the effects of hepatic fatty acid transporter 5 (FATP5) gene expression on the levels of free long-chain fatty acid (LCFA) in mice with metabolic associated fatty liver disease (MAFLD).Methods:From June to December 2022, a prospective study was conducted with three experimental groups: wild-type (WT) group, FATP5 gene expression negative (FATP5 -) group, and human FATP5 gene expression positive (hFATP5 +) group, with 10 mice in each group. Each group of mice was fed a high-fat diet for 16 weeks to establish a model of MAFLD. Hepatic tissue changes were observed using hematoxylin-eosin staining. The liver mass and liver coefficient of the mice were measured. Total cholesterol (TC), triglycerides (TG), aspartate aminotransferase (AST), alanine aminotransferase (ALT), uric acid (UA), and LCFA levels were determined using an automatic biochemical analyzer. Blood glucose (Glu) levels were measured using a blood glucose analyzer. The liver mass and liver coefficient, TC and TG levels, AST and ALT levels, Glu and UA levels, and LCFA levels were compared among the three groups. Results:In the WT group, there was significant inflammatory cell infiltration within the hepatocytes and a large amount of fat accumulation. In the FATP5 - group, the inflammatory cell infiltration in the hepatocytes was mild with slight fat accumulation. In the hFATP5 + group, the inflammatory cell infiltration in the hepatocytes was severe, with great fat accumulation. The liver mass [(1.27 ± 0.25) g], liver coefficient (2.38 ± 0.19), TC [(1.82 ± 0.26) mmol/L], TG [(0.93 ± 0.24) mmol/L], AST [(169.95 ± 37.73) U/L], ALT [(95.36 ± 21.49) U/L], Glu [(8.34 ± 1.52) mmol/L], and UA [(74.32 ± 15.52) μmol/L] in the FATP5 - group were all significantly lower than those in the WT group [(1.61 ± 0.23) g, (2.71 ± 0.20), (2.31 ± 0.28) mmol/L, (1.34 ± 0.21) mmol/L, (278.31 ± 43.24) U/L, (147.32 ± 28.81) U/L, (10.52 ± 1.24) mmol/L, (96.28 ± 17.43) μmol/L], while the LCFA level [(3.57 ± 0.48) mg/L] in the FATP5 - group was significantly higher than that in the WT group [(2.63 ± 0.56) mg/L] ( t = 3.17, 3.78, 4.06, 4.07, 5.97, 4.57, 3.51, 2.98, 4.03, all P < 0.05). In the hFATP5 + group, the liver mass [(1.92 ± 0.30) g], liver coefficient (2.95 ± 0.23), TC [(2.59 ± 0.24) mmol/L], TG [(1.76 ± 0.35) mmol/L], AST [(341.22 ± 48.98) U/L], ALT [(189.45 ± 17.97) U/L], Glu [(13.21 ± 1.98) mmol/L], and UA [(117.74 ± 18.38) μmol/L] were all significantly higher than those in the WT group, while the LCFA level [(3.57 ± 0.48) mg/L] in the FATP5 + group was significantly lower than that in the WT group ( t = 2.59, 2.49, 2.40, 3.25, 3.04, 3.92, 3.64, 2.68, 3.19, all P < 0.05). Conclusions:The absence of FATP5 in the liver can elevate blood levels of LCFA in mice with MAFLD, reduce food intake, and help alleviate the symptoms of MAFLD.

Objective To investigate the expressions of IL-18,IL-18 binding protein isoform a(IL-18BPa)and IL-18Rα in blood CD4+Th1 cells of patients with allergic asthma(AA),and with allergic rhinitis and asthma syndrome(ARA),and the effects of allergens on their expressions.Methods Blood samples were collected from patients with AA,ARA and healthy control(HC)subjects.Flow cytometry was used to evaluate the actions of allergens on the expressions of IL-18,IL-18BPa and IL-18Rα in CD4+Th1 cells.Ovalbumin(OVA)-induced AA mouse models were established,and the expression level of IL-18Rα along with the influence of IL-18 on its expression in blood and lung Th1 cells was also explored by flow cytometry.Results Compared with HC,we observed increased IL-18 while decreased IL-18BPa expression in blood Th1 cells of AA and ARA patients.Moreover,allergens upregulated the expression levels of IL-18,IL-18BPa and IL-18Rα in Th1 cells of patients with AA and ARA.Additionally,intratracheal challenge with OVA plus IL-18 increased the proportions of blood and lung Th1 cells of HC and AA mice,and upregulated IL-18Rα expression on blood and lung Th1 cells of HC mice.Conclusion Allergens may be involved in the pathogenesis of airway allergic diseases by inducing the expressions of IL-18 and IL-18Rα in CD4+Th1 cells.